Part:BBa_K5527002

amilCP yellow, Yellow chromoprotein

This part is the coding sequence of a yellow chromoprotein. The part was made by a substitution at position 157, which in the original amilCP (BBa_K592009) is asparagine, to glutamic acid.

Usage and Biology

amilCP is a chromoprotein from the coral Acropora millepora, which can give it a blue color. It belongs to a larger group of GFP-like proteins, but unlike GFP it isn’t fluorescent. It occurs as a homodimer, where each monomer where each monomer makes up a beta-barrel consisting of 11 beta-strands. The chromophore, made up of Gln62, Tyr63 and Gly64, is connected to an alpha-helix running through the barrel (Ahmed et al. 2022).

This part can be used as a reporter gene. In contrast to the common fluorescent reporters such as GFP, this part absorbs light of the visible spectra. This means no special equipment is needed to detect activity, and the cells don’t have to be exposed to UV light, which could harm them (Liljeruhm et al. 2018).

Characterization

Figure 1: Wild type amilCP, purple, light purple and yellow mutants in liquid culture.

Figure 2: Pellets of wild type amilCP, purple, light purple and yellow mutants.

Figure 3: Pictures of LB-agar plates with different amilCP mutants. Top left: purple mutant. Top right: wild type amilCP. Bottom left: yellow mutant (this part). Bottom right: light purple mutant.

Figure 4: Absorbance spectrum. Maximum at 415 nm.

References

Ahmed FH, Caputo AT, French NG, Peat TS, Whitfield J, Warden AC, Newman J, Scott C. 2022. Over the rainbow: structural characterization of the chromoproteins gfasPurple, amilCP, spisPink and eforRed. Acta Crystallographica Section D, Structural Biology 78: 599–612.

Liljeruhm J, Funk SK, Tietscher S, Edlund AD, Jamal S, Wistrand-Yuen P, Dyrhage K, Gynnå A, Ivermark K, Lövgren J, Törnblom V, Virtanen A, Lundin ER, Wistrand-Yuen E, Forster AC. 2018. Engineering a palette of eukaryotic chromoproteins for bacterial synthetic biology. Journal of Biological Engineering 12: 8.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]